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Molten globule as an intermediate on the human prostatic phosphatase folding pathway
1Institute of Medical Biochemistry, Collegium Medicum, Jagiellonian University, Cracow, Poland.
Acta Biochimica Polonica
|January 1, 1997
Summary
Human prostatic acid phosphatase (hPAP) refolding involves a molten globule intermediate. This intermediate, characterized by a relaxed beta-structure, interacts with hydrophobic probes and lipids, influencing the final protein structure.
Area of Science:
- Biochemistry
- Protein Folding
- Enzymology
Background:
- Human prostatic acid phosphatase (hPAP) is a secretory homodimeric enzyme.
- Understanding protein refolding is crucial for enzyme function and stability.
Purpose of the Study:
- To investigate the kinetic intermediate during hPAP refolding.
- To characterize the structural properties of the refolding intermediate.
- To examine the influence of lipids on hPAP refolding.
Main Methods:
- Protein denaturation and refolding using urea and dilution.
- Circular dichroism and intrinsic fluorescence spectroscopy.
- Chromatographic analysis and dye-binding assays (1-anilinonaphthalene-8-sulfonate, Congo Red).
- Interaction studies with phosphatidylserine liposomes.
Main Results:
- Refolding experiments confirmed a monomeric molten globule intermediate with a relaxed beta-structure.
- Increased binding of hydrophobic probes and Congo Red supported the molten globule state.
- Liposomes perturbed the refolding process, leading to intermediate binding or aggregate formation.
Conclusions:
- The refolding of hPAP proceeds via a molten globule intermediate.
- Lipid interactions can significantly alter protein refolding pathways and outcomes.